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目的:以6-羟基多巴胺(6-OHDA)作用于大鼠肾上腺嗜铬细胞瘤细胞(PC12cells)以诱导其凋亡,然后在其中分别加入神经生长因子(NGF)及c-Jun氨基端激酶(JNK)阻断剂SP600125,研究在加入NGF后JNK的活性与凋亡的关系。方法:实验分为对照组、6-OHDA组、NGF组、6-OHDA+NGF组、6-OHDA+JNK阻断剂SP600125组,以流式细胞分析法检测各组PC12细胞的凋亡率,以免疫印迹(Westernblotting)法检测各组PC12细胞JNK的活化情况。结果:6-OHDA导致PC12细胞凋亡,JNK1活性提高;预孵SP600125或NGF15min后再加入6-OHDA则PC12细胞凋亡率及JNK1活性均降低。结论:JNK1参与了6-OHDA致PC12细胞凋亡作用,NGF抗6-OHDA所诱导的PC12细胞凋亡作用与其抑制JNK的活化有关。
OBJECTIVE: To induce apoptosis of rat adrenal pheochromocytoma cells (PC12cells) by 6-hydroxydopamine (6-OHDA), then add nerve growth factor (NGF) and c-Jun N-terminal kinase JNK) blocker SP600125 to study the relationship between JNK activity and apoptosis after addition of NGF. Methods: The experiment was divided into control group, 6-OHDA group, NGF group, 6-OHDA + NGF group and 6-OHDA + JNK blocker SP600125 group. Flow cytometry was used to detect the apoptosis rate of PC12 cells, The activation of JNK in each group of PC12 cells was detected by Western blotting. Results: 6-OHDA induced apoptosis of PC12 cells and increased JNK1 activity. After preincubated with SP600125 or NGF for 15 minutes, the apoptosis rate of PC12 cells and the activity of JNK1 decreased after adding 6-OHDA. CONCLUSION: JNK1 is involved in the apoptosis of PC12 cells induced by 6-OHDA. The apoptosis of PC12 cells induced by NGF and 6-OHDA is related to the inhibition of JNK activation.